Evaluation of probiotic potential of yeasts isolated from traditional cheeses manufactured in Serbia and Croatia

The aim of this study was to investigate the in vitro probiotic potential of dairy yeast isolates from artisanal cheeses manufactured in Serbia and Croatia. Methods. Twelve yeast strains isolated in from artisanal fresh soft and white brined cheeses manufactured in Serbia and Croatia were used in the study. Survival in chemically-simulated gastrointestinal conditions, adherence to epithelial intestinal cells and proliferation of gut-associated lymphoid tissue (GALT) cells were evaluated. Results. The results revealed that two strains of Kluyvereomyces lactis ZIM 2408 and ZIM 2453 grew above one log unit ( and #916; log CFU/ml) in the complex colonic medium during 24 h of cultivation, while Torulaspora delbrueckii ZIM 2460 was the most resistant isolate in chemically-simulated conditions of gastric juice and upper intestinal tract. It was demonstrated that the strains Kluyvereomyces lactis ZIM 2408 and ZIM2441 and Saccharomyces cerevisiae ZIM 2415 were highly adhesive to Caco-2 cells, while strains Kluyvereomyces lactis ZIM 2408 and Debaryomyces hansenii ZIM 2415 exhibit the highest adhesion percentage to HT29-MTX cells. All strains significantly (p lt 0.0001) decreased the proliferation of gut-associated lymphoid tissue (GALT) cells suggesting the possible strain-specific immunomodulatory potential of the isolates. Conclusion. The dairy yeast isolates exhibit the strain-specific probiotic properties. Particularly, the strain K. lactis ZIM 2408 appears to be the best probiotic candidate in terms of all three criteria. Taking into account their immunomodulatory potential, the yeast isolates could be further tested for specific probiotic applications and eventually included in functional food formulated for patients suffering from diseases associated with an increased inflammatory status.[http://www.jocmr.com/fulltext/55-1410953223.pdf?1669198006

Evaluation of autochthonous lactic acid bacteria as starter cultures for production of white pickled and fresh soft cheeses

In order to preserve the traditional manufacturing of white pickled (WPC) and fresh soft cheeses (FSC), well-characterized autochthonous lactic acid bacteria (LAB) with advantageous characteristics were applied for the production of the cheeses at small industrial scale under the controlled conditions. Selected LAB for design of defined mixed starter cultures belonged to Lactococcus lactis ZGBP5-9, Enterococcus faecium ZGPR1-54 and Lactobacillus plantarum ZGPR2-25 for FSC production and to Lc. lactis BGAL1-4, Lactobacillus brevis BGGO7-28 and Lb. plantarum BGGO7-29 for WPC production. A sensory evaluation indicated that the cheeses obtained by inoculation with selected autochthonous LAB are similar to the traditional cheese and received the best scores. Viable cell counts of LAB used for the production of both type chesses was high, over 10(6) cfu g(-1). High viability of the surveyed strains was supported with PCR-DGGE, which confirm the retention of selected LAB strains as starter cultures in cheese production. Next, PFGE analysis showed that each single strains, selected in particular cheese mixed culture, revealed unique SmaI PFGE pattern that could enable efficient discrimination and monitoring of the strains in industrial process. As some of the selected LAB strains are attributed as potential probiotics, produced cheeses could be considered as functional food

Characterisation of the yeast and mould biota in traditional white pickled cheeses by culture-dependent and independent molecular techniques

Artisanal white pickled cheese of Western Serbia is a product of complex microbial community which detection by culture-dependent method only is hampered by its limitations. Thus, in the present study, we used a culture-independent, semi-quantitative technique based on construction of an internal transcribed spacer (ITS)-clone library from metagenomic DNA. This approach, based on direct DNA extraction followed by amplification of fungal internal transcribed regions (ITS) cloned into plasmid and restricted by endonucleases, revealed greater species richness in analysed cheeses and their by-products (17 species in total) compared to the more commonly used techniques of the culture-dependent method (8 species) and LSU-DGGE (10 species). The most frequently occurring yeast species which are commonly associated with cheeses production were Debaryomyces hansenii, Kluyveromyces lactis and Candida zeylanoides. On the other hand, Yarrowia lipolytica and Galactomyces geotrichum were detected only in one cheese sample. Moreover, some species, mainly moulds (Filobasidium globisporum, Cladosporium sp., Aspergillus sp. or Alternaria sp.) were identified only by culture-independent methods. The discrepancies between the techniques were confirmed by low correlation factor and by different indices of general biodiversity and dominance of species. The ITS-clone library approach provides the opportunity to analyse complex fungal communities associated with food products

Evaluation of lactic acid bacteria and yeast diversity in traditional white pickled and fresh soft cheeses from the mountain regions of Serbia and lowland regions of Croatia

The goal of this study was the characterisation of indigenous lactic acid bacteria (LAB) and yeasts isolated from nine white pickled (BG) and nine fresh soft (ZG) artisanal cheeses collected in Serbia and Croatia. While LAB were present in all of the cheeses collected, yeasts were found in all BG cheeses but only in three ZG cheese samples. High LAB and yeast species diversity was determined (average H-L' = 0.4 and H-Y' = 0.8, respectively). The predominant LAB species in white pickled (BC) cheeses were Lactococcus lactis, Lactobacillus plantarum, and Leuconostoc mesenteroides, while in fresh soft (ZG) cheeses the most dominant LAB species were L. lactis, Enterococcus faecalis, and Leuconostoc pseudomesenteroides. Among the 20 yeast species found, Debaryomyces hansenii, Candida zeylanoides, and Torulaspora delbrueckii were found to be predominant in BC cheeses, while Yarrowia lipolytica was predominant in ZG cheeses. The characterisation of metabolic and technological potentials revealed that 53.4% of LAB isolates produced antimicrobial compounds, 44.3% of LAB strains showed proteolytic activity, while most of the yeast species possessed either lipolytic or proteolytic activity. In conclusion, the results obtained in this study showed that the composition of LAB and yeast populations in white pickled and fresh soft cheeses is region specific. The knowledge gained in this study could eventually be used to select region specific LAB and yeast strains for the production of white pickled and fresh soft artisanal cheeses with geographically specific origins under controlled conditions

Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese

The aim of this study is to gain insight into the probiotic potential of autochthonous lactic acid bacteria (LAB) isolated from artisanal fresh soft and white pickled cheeses. Eleven out of 86 LAB isolates from traditionally produced artisanal fresh soft and white pickled cheeses which survived the most rigorous simulated gastrointestinal tract conditions and did not show resistance to antibiotics were subjected to further evaluation for functional probiotic properties. The ability of the examined strains to assimilate cholesterol in the presence of bile salts was strain dependent, with the highest percentage of cholesterol assimilated by strain Lactobacillus brevis BGGO7-28 possessing S-layer proteins on its cell surface. The growth of strains with mannitol or lactulose as the only carbon source was better than with fructooligosaccharides (FOS) and inulin as prebiotic substrates, which should be considered in the production of synbiotics. Moreover, the results demonstrated that the strains were highly adhesive to human enterocyte-like Caco-2 cells and to a lesser extent to HT29-MTX cells, with the exception of strain Lb. brevis BGGO7-28, which showed similar percentage of adhesion to both cell lines. This strain was the only one with the acidic cell surface, while other examined strains have the cell surfaces with electron donor and basic properties. In addition, all selected strains decreased the proliferation of gut-associated lymphoid tissue (GALT) cells, suggesting possible immunomodulatory potential of the isolates. Finally, the number of viable cells in dry active preparations after lyophilisation depended on the lyoprotectant used (inulin, FOS or skimmed milk), as well as on the strain subjected to lyophilisation. In conclusion, the results obtained in this study demonstrate that particular dairy LAB isolates exhibit strain-specific probiotic properties. Thus, they could be further examined as part of mixed autochthonous starter cultures for traditional cheese production under controlled conditions

Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese

The aim of this study is to gain insight into the probiotic potential of autochthonous lactic acid bacteria (LAB) isolated from artisanal fresh soft and white pickled cheeses. Eleven out of 86 LAB isolates from traditionally produced artisanal fresh soft and white pickled cheeses which survived the most rigorous simulated gastrointestinal tract conditions and did not show resistance to antibiotics were subjected to further evaluation for functional probiotic properties. The ability of the examined strains to assimilate cholesterol in the presence of bile salts was strain dependent, with the highest percentage of cholesterol assimilated by strain Lactobacillus brevis BGGO7-28 possessing S-layer proteins on its cell surface. The growth of strains with mannitol or lactulose as the only carbon source was better than with fructooligosaccharides (FOS) and inulin as prebiotic substrates, which should be considered in the production of synbiotics. Moreover, the results demonstrated that the strains were highly adhesive to human enterocyte-like Caco-2 cells and to a lesser extent to HT29-MTX cells, with the exception of strain Lb. brevis BGGO7-28, which showed similar percentage of adhesion to both cell lines. This strain was the only one with the acidic cell surface, while other examined strains have the cell surfaces with electron donor and basic properties. In addition, all selected strains decreased the proliferation of gut-associated lymphoid tissue (GALT) cells, suggesting possible immunomodulatory potential of the isolates. Finally, the number of viable cells in dry active preparations after lyophilisation depended on the lyoprotectant used (inulin, FOS or skimmed milk), as well as on the strain subjected to lyophilisation. In conclusion, the results obtained in this study demonstrate that particular dairy LAB isolates exhibit strain-specific probiotic properties. Thus, they could be further examined as part of mixed autochthonous starter cultures for traditional cheese production under controlled conditions